Very late antigen-4 (VLA-4) is a transmembrane integrin protein that is highly expressed in aggressive forms of metastatic melanoma. A small-molecule peptidomimetic, LLP2A, was found to have a low pM affinity binding to VLA-4. Because LLP2A itself does not inhibit cancer cell proliferation and survival, it is an ideal candidate for the imaging and delivery of therapeutic payloads. An analog of [¹⁷⁷Lu]Lu-labeled-LLP2A was previously investigated as a therapeutic agent in melanoma tumor-bearing mice, resulting in only a modest improvement in tumor growth inhibition, likely due to rapid clearance of the agent from the tumor. To improve the pharmacokinetic profile, DOTAGA-PEG₄-LLP2A with a 4-(p-iodophenyl)butyric acid (pIBA) albumin binding moiety was synthesized. We demonstrate the feasibility of this albumin binding strategy by comparing in vitro cell binding assays and in vivo biodistribution performance of [¹⁷⁷Lu]Lu-DOTAGA-PEG4-LLP2A ([¹⁷⁷Lu]Lu-1) to the albumin binding [¹⁷⁷Lu]Lu-DOTAGA-pIBA-PEG₄-LLP2A ([¹⁷⁷Lu]Lu-2). In vitro cell binding assay results for [¹⁷⁷Lu]Lu-1 and [¹⁷⁷Lu]Lu-2 showed K_d values of 0.40 ± 0.07 and 1.75 ± 0.40 nM, with similar B_max values of 200 ± 6 and 315 ± 15 fmol/mg, respectively. In vivo biodistribution data for both tracers exhibited specific uptake in the tumor, spleen, thymus, and bone due to endogenous expression of VLA-4. Compound [¹⁷⁷Lu]Lu-2 exhibited a much longer blood circulation time compared to [¹⁷⁷Lu]Lu-1. The tumor uptake for [¹⁷⁷Lu]Lu-1 was highest at 1 h (∼15%ID/g) and that for [¹⁷⁷Lu]Lu-2 was highest at 4 h (∼23%ID/g). Significant clearance of [¹⁷⁷Lu]Lu-1 from the tumor occurs at 24 h (<5%ID/g) while[¹⁷⁷Lu]Lu-2 is retained for greater than 96 h (∼10%ID/g). An efficacy study showed that melanoma tumor-bearing mice receiving compound [¹⁷⁷Lu]Lu-2 given in two fractions (2 × 14.8 MBq, 14 days apart) had a greater median survival time than mice administered a single 29.6 MBq dose of compound [¹⁷⁷Lu]Lu-1, while a single 29.6 MBq dose of [¹⁷⁷Lu]Lu-2 imparted hematopoietic toxicity. The in vitro and in vivo data show addition of pIBA to [¹⁷⁷Lu]Lu-DOTAGA-PEG₄-LLP2A slows blood clearance for a higher tumor uptake, and there is potential of [¹⁷⁷Lu]Lu-2 as a theranostic in fractionated administered doses.

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